LOCUS       sequence-502042-        8898 bp    DNA     circular SYN 06-MAR-2026
DEFINITION  mammalian expression of human RPA70 N-terminally fused to mVenus and
            RPA32 and RPA14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8898)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 8898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 6, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pmVen-RPA70-IRES-RPA32-IRES-RPA14
FEATURES             Location/Qualifiers
     source          1..8898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      632..641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(638..640,641..643,644..1354)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(683..704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(944..963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1291..1312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5711..5832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5748..5767)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5802..5821
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5839..6294)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5976..5997)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6188..6207
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6321..6425
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6427..6784
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6427..6618
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6450..6470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6635..6770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6819..7613
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6819..7613
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6873..6892)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7483..7502
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7845..7892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8221..8809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8710..8729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
      661 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      721 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
      781 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
      841 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      901 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
      961 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1021 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1081 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     1141 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     1201 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct
     1261 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1321 cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc gagctcgtat
     1381 ggtcggccag ctgagcgagg gggccattgc ggccatcatg cagaaggggg atacaaacat
     1441 aaagcccatc ctccaagtca tcaacatccg tcccattact acggggaata gtccgccgcg
     1501 ttatcgactg ctcatgagtg atggattgaa cactctatcc tctttcatgt tggcgacaca
     1561 gttgaaccct ctcgtggagg aagaacaatt gtccagcaac tgtgtatgcc agattcacag
     1621 atttattgtg aacactctga aagacggaag gagagtagtt atcttgatgg aattagaagt
     1681 tttgaagtca gctgaagcag ttggagtgaa gattggcaat ccagtgccct ataatgaagg
     1741 actcgggcag ccgcaagtag ctcctccagc gccagcagcc agcccagcag caagcagcag
     1801 gccccagccg cagaatggaa gctcgggaat gggttctact gtttctaagg cttatggtgc
     1861 ttcaaagaca tttggaaaag ctgcaggtcc cagcctgtca cacacttctg ggggaacaca
     1921 gtccaaagtg gtgcccattg ccagcctcac tccttaccag tccaagtgga ccatttgtgc
     1981 tcgtgttacc aacaaaagtc agatccgtac ctggagcaac tcccgagggg aagggaagct
     2041 tttctcccta gaactggttg acgaaagtgg tgaaatccga gctacagctt tcaatgagca
     2101 agtggacaag ttctttcctc ttattgaagt gaacaaggtg tattatttct cgaaaggcac
     2161 cctgaagatt gctaacaagc agttcacagc tgttaaaaat gactacgaga tgaccttcaa
     2221 taacgagact tccgtcatgc cctgtgagga cgaccatcat ttacctacgg ttcagtttga
     2281 tttcacgggg attgatgacc tcgagaacaa gtcgaaagac tcacttgtag acatcatcgg
     2341 gatctgcaag agctatgaag acgccactaa aatcacagtg aggtctaaca acagagaagt
     2401 tgccaagagg aatatctact tgatggacac atctgggaag gtggtgactg ctacactgtg
     2461 gggggaagat gctgataaat ttgatggttc tagacagccc gtgttggcta tcaaaggagc
     2521 ccgagtctct gatttcggtg gacggagcct ctccgtgctg tcttcaagca ctatcattgc
     2581 gaatcctgac atcccagagg cctataagct tcgtggatgg tttgacgcag aaggacaagc
     2641 cttagatggt gtttccatct ctgatctaaa gagcggcgga gtcggaggga gtaacaccaa
     2701 ctggaaaacc ttgtatgagg tcaaatccga gaacctgggc caaggcgaca agccggacta
     2761 ctttagttct gtggccacag tggtgtatct tcgcaaagag aactgcatgt accaagcctg
     2821 cccgactcag gactgcaata agaaagtgat tgatcaacag aatggattgt accgctgtga
     2881 gaagtgcgac accgaatttc ccaatttcaa gtaccgcatg atcctgtcag taaatattgc
     2941 agattttcaa gagaatcagt gggtgacttg tttccaggag tctgctgaag ctatccttgg
     3001 acaaaatgct gcttatcttg gggaattaaa agacaagaat gaacaggcat ttgaagaagt
     3061 tttccagaat gccaacttcc gatctttcat attcagagtc agggtcaaag tggagaccta
     3121 caacgacgag tctcgaatta aggccactgt gatggacgtg aagcccgtgg actacagaga
     3181 gtatggccga aggctggtca tgagcatcag gagaagtgca ttgatgtgag gatctttcgc
     3241 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     3301 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     3361 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     3421 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     3481 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     3541 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     3601 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     3661 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     3721 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     3781 gacgtggttt tcctttgaaa aacacgataa gaattctgcc accatgtgga acagtggatt
     3841 cgaaagctat ggcagctcct catacggggg agccggcggc tacacgcagt ccccgggggg
     3901 ctttggatcg cccgcacctt ctcaagccga aaagaaatca agagcccgag cccagcacat
     3961 tgtgccctgt actatatctc agctgctttc tgccactttg gttgatgaag tgttcagaat
     4021 tgggaatgtt gagatttcac aggtcactat tgtggggatc atcagacatg cagagaaggc
     4081 tccaaccaac attgtttaca aaatagatga catgacagct gcacccatgg acgttcgcca
     4141 gtgggttgac acagatgaca ccagcagtga aaacactgtg gttcctccag aaacatatgt
     4201 gaaagtggca ggccacctga gatcttttca gaacaaaaag agcctggtag cctttaagat
     4261 catgcccctg gaggatatga atgagttcac cacacatatt ctggaagtga tcaatgcaca
     4321 catggtacta agcaaagcca acagccagcc ctcagcaggg agagcaccta tcagcaatcc
     4381 aggaatgagt gaagcaggga actttggtgg gaatagcttc atgccagcaa atggcctcac
     4441 tgtggcccaa aaccaggtgt tgaatttgat taaggcttgt ccaagacctg aagggttgaa
     4501 ctttcaggat ctcaagaacc agctgaaaca catgtctgta tcctcaatca agcaagctgt
     4561 ggattttctg agcaatgagg ggcacatcta ttctactgtg gatgatgacc attttaaatc
     4621 cacagatgca gaataaggat ccaattccgc ccctctccct cccccccccc taacgttact
     4681 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     4741 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     4801 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     4861 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     4921 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     4981 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     5041 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     5101 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     5161 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgataa
     5221 agatctcatg gtggacatga tggacttgcc caggtcgcgc atcaacgccg gcatgctagc
     5281 tcaattcatc gacaagcctg tctgcttcgt agggaggctg gaaaagattc atcccaccgg
     5341 aaaaatgttt attctttcag atggagaagg aaaaaatgga accatcgagt tgatggaacc
     5401 ccttgatgaa gaaatctctg gaattgtgga agtggttgga agagtaaccg ccaaggccac
     5461 catcttgtgt acatcttatg tccagtttaa agaagatagc catccttttg atcttggact
     5521 ttacaatgaa gctgtgaaaa ttatccatga cttccctcag ttttatcctt tagggattgt
     5581 gcaacatgat tgatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     5641 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     5701 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5761 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5821 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     5881 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     5941 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     6001 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     6061 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     6121 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     6181 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     6241 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     6301 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6361 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6421 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     6481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6601 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6661 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6721 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6781 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     6841 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     6901 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     6961 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     7021 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     7081 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     7141 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     7201 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     7261 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     7321 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     7381 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     7441 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     7501 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     7561 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     7621 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     7681 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     7741 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     7801 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     7861 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     7921 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     7981 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     8041 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     8101 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8161 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8221 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8281 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8341 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8401 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8461 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8521 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8581 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     8641 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8701 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8761 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8821 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8881 gttatcccct gattctgt
//
